title: A new approach for diagnosis of bovine coronavirus using a reverse transcription recombinase polymerase amplification assay.
abstract: Bovine coronavirus (BCoV) is an economically significant cause of calf scours and winter dysentery of adult cattle, and may induce respiratory tract infections in cattle of all ages. Early diagnosis of BCoV helps to diminish its burden on the dairy and beef industry. Real-time RT-PCR assay for the detection of BCoV has been described, but it is relatively expensive, requires well-equipped laboratories and is not suitable for on-site screening. A novel assay, using reverse transcription recombinase polymerase amplification (RT-RPA), for the detection of BCoV is developed. The BCoV RT-RPA was rapid (10-20 min) and has an analytical sensitivity of 19 molecules. No cross-reactivity with other viruses causing bovine gastrointestinal and/or respiratory infections was observed. The assay performance on clinical samples was validated by testing 16 fecal and 14 nasal swab specimens and compared to real-time RT-PCR. Both assays provided comparable results. The RT-RPA assay was significantly more rapid than the real-time RT-PCR assay. The BCoV RT-RPA constitutes a suitable accurate, sensitive and rapid alternative to the common measures used for BCoV diagnosis. In addition, the use of a portable fluorescence reading device extends its application potential to use in the field and point-of-care diagnosis.
authors.affiliation: Department of Botany and Microbiology, College of Science, King Saud University, 11451 Riyadh, Saudi Arabia; Department of Virology, Faculty of Veterinary Medicine, Cairo University, 12211 Giza, Egypt.
authors.forename: H M
authors.initials: HM
authors.lastname: Amer
authors.forename: A
authors.initials: A
authors.lastname: Abd El Wahed
authors.forename: M A
authors.initials: MA
authors.lastname: Shalaby
authors.forename: F N
authors.initials: FN
authors.lastname: Almajhdi
authors.forename: F T
authors.initials: FT
authors.lastname: Hufert
authors.forename: M
authors.initials: M
authors.lastname: Weidmann
mesh_terms: D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000821:virology; D018352:Coronavirus Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D017938:Coronavirus, Bovine / Q000235:genetics / Q000302:isolation & purification*; D005243:Feces / Q000821:virology; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009296:Nasal Cavity / Q000821:virology; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D019095:Point-of-Care Systems; D045522:Recombinases / Q000378:metabolism*; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D013997:Time Factors; D014730:Veterinary Medicine / Q000379:methods; D014773:Virology / Q000379:methods
chemical_list: D045522:Recombinases
keywords: BCoV; Bovine coronavirus; LAMP; MEM; Molecular diagnostic techniques; PCR; Point-of-care systems; RPA; RT; bovine coronavirus; loop-mediated isothermal amplification; minimal essential medium; polymerase chain reaction; recombinase polymerase amplification; reverse transcriptase